Turn To Gl Fusion Protein Gene Of Rice Gene Expression Product Allergenic And Acute Toxicity Study

Genetically modified food allergens and toxic security issue has been a growing concern, is an important aspect of food safety issues. Especialy, as the major food in the world, the transgenic rice have aroused more and more fouces. At the national funding of new varieties of genetically modified organism cultivate major projects (2009ZX-08011-003B), digestion of this study refer to the WHO/FAO genetically modified food allergens evaluation strategy by the amino acid sequence similarity, the GL transgenic rice totel protein stability studies, use the digestion stability and acute toxicity studies to evaluat the safety of GL fusion protein from transgenic rice.Reference to national standards of the People’s Republic of China, according to the total protein concentration gradient of5g/L,25g/L of simulated gastric digestion, and total protein concentration of2g/L,10g/L in simulated intestinal digestion, GL fusion protein gene transgenic rice protein digestion stability. The results showed that the gastric juice5g/L can be completely degraded in2min,25g/Lof digestion, utill to60min there observed degradation fragment; in intestinal juice digastion,2g/L,10g/L, total protein degraded in the15s completely. The first two-step method simulated digestion after gastric juice digestion1h, and then go on the intestinal digestion, the results showed that gastric juice the role of1h25g/L protein degradation fragment was completely degraded by SIF for5min. Transgenic rice protein’s concentration equivalent to the standard’s five times, after one hour of the gastric juice incubation, in intestinal fluid for5min completely degraded, the static simulation of digestion with two-step method is more in line with the human digestive tract to digest and can be used to evaluat the digestion stability.Use gel recycling GL fusion protein digestion stability and acute toxicity studies, preliminary evaluation of the security of GL fusion protein. The simulated gastric/intestinal digestion results analysis by SDS-PAGE electrophoresis and Western blot; intravenous injection of GL fusion protein to observe the acute toxicity with ICR mice. the results show that the GL fusion protein in simulated gastric digestion for10min to degrade completely, in simulated intestinal digestion complete decomposition for0s; acute toxicity experiments showed that the growth of the body weight of mice have no significance difference, organ coefficient and biochemical indicators of protein group compared with the control group, there is no physiological and biochemical significance of the differences, and within the normal range. GL fusion protein in a simulated gastrointestinal fluid digestion experiments with no stability, and did not have a negative impact in the mice.GL fusion protein amino acid sequence compared with the current internationally recognized eight major known allergen categories’s (peanut, soy, milk, eggs, fish, shellfish, cereals and tree nuts) amino acid sequences, no more than35%similarity sequence, nor any6consecutive identical amino acids. Description of the GL fusion protein with known allergens are no similar amino acid sequence; unstability of exogenous protein digestion in simulated gastrointestinal fluid digestion experiments, and the mice did not have a negative impact.Preliminarily determined the security of the GL fusion protein in transgenic rice.